The role of maladaptive VEGFR2 signaling in renal stroma for chronic kidney disease

肾间质中适应不良的 VEGFR2 信号传导在慢性肾脏病中的作用

基本信息

批准号：
10739504
负责人：
Takuto Chiba
金额：
$ 12.14万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-24 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10739504
关键词：
Acute Renal Failure with Renal Papillary Necrosis Age Attenuated Blood Vessels Cell Separation Cells Chronic Kidney Failure Cisplatin Coculture Techniques Critical Illness Data Dependovirus Disease Disease Progression Disease model Endothelial Cells Endothelium Expenditure Faculty Fatty Acids Female Genes Heart Arrest Heterogeneity Home Human In Situ Hybridization In Vitro Inflammation Inflammatory Injury Injury to Kidney KDR gene Kidney Kidney Diseases Knock-out Knockout Mice Mediating Medicare Medicine Mentored Research Scientist Development Award Mentors Mesenchymal Methods Mitochondria Modeling Molecular Mus Myofibroblast Nature PDH kinase PTPRJ gene Pathway interactions Patients Pericytes Phenotype Platelet-Derived Growth Factor Play Population Predictive Factor Process Production Reperfusion Injury Reporter Research Respiration Risk Role Signal Transduction Small Interfering RNA Source Tamoxifen Testing Therapeutic Thrombospondin 1 Universities Validation Volatile Fatty Acids adeno-associated viral vector cell injury chemotherapeutic agent etomoxir experimental study fatty acid metabolism fatty acid oxidation inhibitor innovation kidney cell loss of function male mouse model pharmacologic preservation programs recruit renal ischemia repaired sex side effect single-cell RNA sequencing therapeutic evaluation therapy development transcriptome sequencing

项目摘要

PROJECT SUMMARY/ ABSTRACT A dire consequence of acute kidney injury (AKI) is a dramatically increased risk to develop chronic kidney disease (CKD). CKD accounts for 6.7% of Medicare expenses. Understanding the mechanisms by which AKI progresses to CKD is essential for developing therapies, for which none currently exist. Renal microvasculature, including pericytes and endothelial cells, are damaged in AKI, leading to recruitment of inflammatory cells which contributes to progression to CKD. However, cellular, and molecular mechanisms that drive this process are largely unknown. Pericytes are a heterogeneous mesenchymal population and have been identified as a major source of myofibroblasts that drives CKD. Understanding the molecular mechanisms that mediates maladaptive endothelial-pericyte crosstalk leading to exacerbated and prolonged inflammation could drive therapeutic exploitation of this phenomenon. Previously, it has been shown that the systemic blockade of Vascular endothelial growth factor receptor 2 (VEGF-R2) blocks CKD progression. My preliminary data, knocking out VegfR2 in renal stromally derived cells (RSC) (termed VegfR2RSC-/-) that includes pericytes, confirms the protective nature of this loss-of-function. I found that, in CKD models by renal ischemia/ reperfusion injury (IRI) as well as by cisplatin, VegfR2RSC-/- mice have attenuated CKD progression, along with having mitigated inflammation and preserved vascular function. My bulk RNA-sequencing analysis with isolated RSCs demonstrates that inflammatory pathways are activated while short-chain fatty acid metabolism pathways are suppressed during AKI-to-CKD transition. Mechanistically, VegfR2RSC-/- kidneys (1) have reduced expression of a pro-inflammatory signaling axis of Thrombospondin-1 (TSP1)/ CD148, and (2) have increased expression of fatty acid metabolism associated genes contributing to the enhanced protection. To home in on the timing of the protection, I have generated a tamoxifen-inducible RSC-specific VegfR2 knockout (iVegfR2RSC-/-) mouse. I found that, after pre-treatment of tamoxifen, iVegfR2RSC-/- mice are significantly protected against AKI. Together, these data informed my overarching hypothesis that renal pericyte-specific VEGF-R2 signaling dysregulates pericyte- endothelial crosstalk stimulating inflammation to exacerbate CKD. I propose the following aims to test this: Aim 1 will test the hypothesis that pericyte-specific VEGF-R2 signaling exacerbates AKI-to-CKD transition. Aim 2 will test the hypothesis that pericyte VEGF-R2 signaling mediates maladaptive pericyte-endothelial crosstalk to exacerbate inflammation, promoting AKI-to-CKD. Aim 3 will test the hypothesis that inhibiting VegfR2 signaling in renal pericytes enhances vascular repair, mitigates inflammation, and blocks progression to CKD after AKI. For the K01 Award, I enlisted innovative mentors. The University of Pittsburgh has an extraordinary number of faculty with research programs focused on AKI, and on vascular medicine. Successful completion of these studies will shed light on how renal pericyte specific VEGF-R2 signaling exacerbates CKD progression.

项目概要/摘要急性肾损伤 (AKI) 的一个可怕后果是患慢性肾病的风险急剧增加（CKD）。 CKD 占医疗保险费用的 6.7%。了解 AKI 进展的机制 CKD 的治疗对于开发治疗方法至关重要，但目前尚无治疗方法。肾脏微血管，包括周细胞和内皮细胞在 AKI 中受损，导致炎症细胞募集，从而导致有助于 CKD 的进展。然而，驱动这一过程的细胞和分子机制是很大程度上不为人知。周细胞是一种异质的间充质群体，已被确定为主要的驱动 CKD 的肌成纤维细胞的来源。了解介导适应不良的分子机制内皮-周细胞串扰导致炎症加剧和延长，可能会推动治疗利用这种现象。此前，已有研究表明，血管的系统性阻断内皮生长因子受体 2 (VEGF-R2) 可阻断 CKD 进展。我的初步数据，淘汰包括周细胞在内的肾间质来源细胞 (RSC)（称为 VegfR2RSC-/-）中的 VegfR2 证实了这种功能丧失的保护性质。我发现，在肾缺血/再灌注损伤 (IRI) 引起的 CKD 模型中与顺铂一样，VegfR2RSC-/- 小鼠可以减弱 CKD 的进展，同时减轻炎症和保留血管功能。我使用分离的 RSC 进行的批量 RNA 测序分析表明炎症途径被激活，而短链脂肪酸代谢途径被激活在 AKI 向 CKD 转变期间受到抑制。从机制上讲，VegfR2RSC-/- 肾脏 (1) 的表达减少 Thrombospondin-1 (TSP1)/CD148 的促炎信号轴，并且 (2) 增加了脂肪酸代谢相关基因有助于增强保护。关注时间为了保护，我已经生成了他莫昔芬诱导的 RSC 特异性 VegfR2 敲除 (iVegfR2RSC-/-) 小鼠。我发现经他莫昔芬预处理后，iVegfR2RSC-/- 小鼠可显着预防 AKI。在一起，这些数据告诉我一个总体假设：肾周细胞特异性 VEGF-R2 信号传导失调周细胞内皮串扰刺激炎症，加剧 CKD。我提出以下目标来测试这一点：目标图 1 将检验周细胞特异性 VEGF-R2 信号传导加剧 AKI 向 CKD 转变的假设。目标2将检验周细胞 VEGF-R2 信号传导介导适应不良的周细胞 - 内皮细胞串扰的假设加剧炎症，促进 AKI 向 CKD 转变。目标 3 将检验抑制 VegfR2 信号传导的假设肾周细胞中的 AKI 增强血管修复、减轻炎症并阻止进展为 CKD。为了获得 K01 奖，我招募了创新导师。匹兹堡大学拥有数量惊人的教授的研究项目主要集中于 AKI 和血管医学。顺利完成这些研究将揭示肾脏周细胞特异性 VEGF-R2 信号传导如何加剧 CKD 进展。